Combination of Doxorubicin and Areca Ethanolic Extract Induces Apoptosis by Increasing Caspase-3 Level on Breast Cancer (T47D) Cells

Fitria Rahmi, Edy Meiyanto, Ratna Asmah Susidarti


Despite causing many side effects, doxorubicin (Dox) is still one of breast cancer drug of choice. Thus, combination of chemotherapy is developed in order to decrease doxorubicin regimen dose. The aim of this research is to examine the combination effect of doxorubicin (dox) and areca extract (AE) on T47D human breast cancer cells. The cytotoxic activity was determined using MTT assay. The combination index (CI) of the combination treatment was calculated to determine the effects (synergistic, additive or antagonistic). The combination application of dox (6-22nM) and AE (8-30µg/ml) on T47D cells showed synergistic (CI<0.9) or additive effect (CI=0.9-1.1). The effective combination of dox-AE was 6 nM - 8 µg/ml on CI<0.5. Apoptosis induction of AE solely and its combination with dox was then observed using double staining method. Moreover, expression of Bax and caspase-3 protein which mediated apoptosis, were observed using immunocytochemistry. Combination of AE and Dox increased expression of Caspase-3 but did not increase expression of Bax. This result showed AE increase the effectiveness of doxorubicin against T47D cells.

Keywords: Breast cancer, doxorubicin, areca extract, T47D cells

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Arai, M., Yoguchi, A., Takizawa, T., Yokoyama, T., Kanda, T., Kurabayashi, M., et al., 2000, Mechanism of Doxorubicin-Induced Inhibition of Sarcoplasmic Reticulum Ca2+-ATPase Gene Transcription, Circ. Res., 86(1), 8-14. Link

Fang, J., Nakamura, H. and Iyer, A.K., 2007, Tumor-targeted Induction of Oxystress for Cancer Theraphy, J. Drug Target, 15(7-8), 475-486. CrossRef

Flusberg, D.A., Numaguchi, Y., Ingber, D.E., 2001, Cooperative Control of Akt Phosphorylation, bcl-2 Expression, and Apoptosis by Cytoskeletal Microfilaments and Microtubules in Capillary Endothelial Cells, Mol. Biol. Cell, 12(10), 3087-3094. Link

Hanahan, D. and Weinberg, R.A., 2011, Hallmarks of Cancer: The Next Generation, Cell, 144(5), 646-674. CrossRef

Itagaki, H., Ohno, T., Hatao, M., Hattori, A., Hayasaka, A., Hayashi, M., et al., 1998, Validation Study on Five Cytotoxicity Assays by JSAAE-VII, Altern. Animal Test. Experiment, 5, 119-130. Link

Jemal, A., Murray, T., Ward, E., Samuels, A., Tiwari, R.C., Ghafoor, A., et al., 2005, Cancer Statistics, 2005, CA Cancer J. Clin., 55(1), 10-30. Link

Korsmeyer, S.J., Wei, M.C., Saito, M., Weiler, S., Oh, K.J. and Schlesinger, P.H., 2000, Pro-Apoptotic Cascade Activates BID, which Oligomerizes Bak or Bax into Pores that Result in the Release of Cytochrome C, Cell Death Differ., 7(12), 1166-1173. CrossRef

Li, X., Lu, Y., Liang, K., Liu, B. and Fan, Z., 2005, Differential Responses to Doxorubicin-Induced Phosphorylation and Activation of Akt in Human Breast Cancer Cells, Breast Cancer Res., 7(5), 589-597. CrossRef

Meiyanto, E., Susidarti, R.A., Handayani, S. and Rahmi, F., 2008, Ekstrak Etanolik Biji Buah Pinang (Areca catechu L.) Mampu Menghambat Proliferasi dan Memacu Apoptosis Sel MCF-7, Indonesian J. Pharm, 19(1), 12-19. Link

Mooney, L.M., Al-Sakkaf, K.A., Brown, B.L. and Dobson, P.R.M., 2002, Apoptotic Mechanisms in T47D and MCF-7 Human Breast Cancer Cells, Br. J. Cancer, 87(8), 909-917. CrossRef

Mullin, A.E. and Claude, B.J., 1998, HER2/neu Oncogene and Sensitivity to the DNA-Interactive Drug Doxorubicin, MJM, 4(1), 9-15. Link

Notarbartolo, M. P., Poma, D., Perri, L., Dusonchet, M. and D’Alessandro, N., 2005, Antitumor Effects of Curcumin, alone or in Combination with Cisplatin or Doxorubicin, on Human Hepatic Cancer Cells. Analysis of Their Possible Relationship to Change in NF-kB Activation Levels and in IAP Gen Expression, Cancer Lett., 224(1), 53-65. CrossRef

Smith, L., Watson, M.B., O'Kane, S.L., Drew, P.J., Lind, M.J. and Cawkwell, L., 2006, The Analysis of Doxorubicin Resistance in Human Breast Cancer Cell Using Antibody Microarray, Mol. Cancer Ther., 5(8), 2115-2120. CrossRef

Thomas, H. and Coley, H.M., 2003, Overcoming Multidrug Resistance in cancer: An Update on the Clinical Strategy of Inhibiting P-Glycoproein, Cancer Control, 10(2), 159-165. CrossRef

Takatani, T., Takahashi, K., Uozumi, Y., Matsuda, T., Ito, T., Schaffer, S.W., et al., 2004, Taurine Prevents the Ischemia-Induced Apoptosis in Cultured Neonatal Rat Cardiomyocytes Through Akt/caspase-9 Pathway, Biochem. Biophys. Res. Commun., 316(2), 484-489. CrossRef

Ueda, J.Y., Tezuka, Y., Banskota., A.H., Tran, Q.L., Tran, Q.K., Harimaya, Y., et al., 2002, Antiproliferative Activity of Vietnamese Medicinal Plants, Biol. Pharm. Bull., 25(6), 753-760. Link

Wetwitayaklung, P. T., Phaechamud, C., Keokitichai, S., and Limmatvapirat, C., 2006, The Study of Antioxidant Capacity in Various parts of Areca catechu L., Naresuen University Journal, 14(1), 1-14. Link


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